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Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro
The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Medknow Publications & Media Pvt Ltd
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955190/ https://www.ncbi.nlm.nih.gov/pubmed/26585695 http://dx.doi.org/10.4103/1008-682X.163189 |
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author | McCabe, Mark J Foo, Caroline FH Dinger, Marcel E Smooker, Peter M Stanton, Peter G |
author_facet | McCabe, Mark J Foo, Caroline FH Dinger, Marcel E Smooker, Peter M Stanton, Peter G |
author_sort | McCabe, Mark J |
collection | PubMed |
description | The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% (P < 0.01), 51% (P < 0.01), and 62% (P < 0.01), respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly (P < 0.01) to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant (P < 0.01) contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function. |
format | Online Article Text |
id | pubmed-4955190 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Medknow Publications & Media Pvt Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-49551902016-07-26 Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro McCabe, Mark J Foo, Caroline FH Dinger, Marcel E Smooker, Peter M Stanton, Peter G Asian J Androl Original Article The Sertoli cell tight junction (TJ) is the key component of the blood-testis barrier, where it sequesters developing germ cells undergoing spermatogenesis within the seminiferous tubules. Hormonally regulated claudin-11 is a critical transmembrane protein involved in barrier function and its murine knockout results in infertility. We aimed to assess quantitatively the significance of the contribution of claudin-11 to TJ function, in vitro, using siRNA-mediated gene silencing. We also conducted an analysis of the contribution of occludin, another intrinsic transmembrane protein of the TJ. Silencing of claudin-11 and/or occludin was conducted using siRNA in an immature rat Sertoli cell culture model. Transepithelial electrical resistance was used to assess quantitatively TJ function throughout the culture. Two days after siRNA treatment, cells were fixed for immunocytochemical localization of junction proteins or lyzed for RT-PCR assessment of mRNA expression. Silencing of claudin-11, occludin, or both resulted in significant decreases in TJ function of 55% (P < 0.01), 51% (P < 0.01), and 62% (P < 0.01), respectively. Data were concomitant with significant decreases in mRNA expression and marked reductions in the localization of targeted proteins to the Sertoli cell TJ. We provide quantitative evidence that claudin-11 contributes significantly (P < 0.01) to Sertoli cell TJ function in vitro. Interestingly, occludin, which is hormonally regulated but not implicated in infertility until late adulthood, is also a significant (P < 0.01) contributor to barrier function. Our data are consistent with in vivo studies that clearly demonstrate a role for these proteins in maintaining normal TJ barrier structure and function. Medknow Publications & Media Pvt Ltd 2016 2015-11-10 /pmc/articles/PMC4955190/ /pubmed/26585695 http://dx.doi.org/10.4103/1008-682X.163189 Text en Copyright: © Asian Journal of Andrology http://creativecommons.org/licenses/by-nc-sa/3.0 This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms. |
spellingShingle | Original Article McCabe, Mark J Foo, Caroline FH Dinger, Marcel E Smooker, Peter M Stanton, Peter G Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title | Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title_full | Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title_fullStr | Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title_full_unstemmed | Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title_short | Claudin-11 and occludin are major contributors to Sertoli cell tight junction function, in vitro |
title_sort | claudin-11 and occludin are major contributors to sertoli cell tight junction function, in vitro |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4955190/ https://www.ncbi.nlm.nih.gov/pubmed/26585695 http://dx.doi.org/10.4103/1008-682X.163189 |
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